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Literature summary extracted from

  • Kaper, T.; Brouns, S.J.; Geerling, A.C.; De Vos, W.M.; Van der Oost, J.
    DNA family shuffling of hyperthermostable beta-glycosidases (2002), Biochem. J., 368, 461-470.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.B26 expression in Escherichia coli Saccharolobus solfataricus
3.2.1.B28 expression in Escherichia coli Pyrococcus furiosus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.B26 additional information generation of functional glycoside hydrolase hybrids by shuffling of the genes coding for Pyrococcus furiosus CelB and Sulfolobus solfataricus LacS. Generating of thermostable hybrid beta-glycosidases and isolating high-performance variants Saccharolobus solfataricus
3.2.1.B28 additional information generation of functional glycoside hydrolase hybrids by shuffling of the genes coding for Pyrococcus furiosus CelB and Sulfolobus solfataricus LacS. Generating of thermostable hybrid beta-glycosidases and isolating high-performance variants Pyrococcus furiosus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.B26 (5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol inhibits activity with 4-nitrophenyl beta-D-galactopyranoside Saccharolobus solfataricus
3.2.1.B26 D-glucose inhibits activity with 4-nitrophenyl beta-D-galactopyranoside Saccharolobus solfataricus
3.2.1.B28 (5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol inhibits activity with 4-nitrophenyl beta-D-galactopyranoside Pyrococcus furiosus
3.2.1.B28 D-glucose inhibits activity with 4-nitrophenyl beta-D-galactopyranoside Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.B26 0.12
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 1.57
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 13.5
-
 cellobiose pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 37.9
-
 lactose pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B28 0.8
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 2.6
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 3.2
-
 cellobiose pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 59.8
-
 lactose pH 5.0, 70°C Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.B26 Saccharolobus solfataricus P22498
-
-
3.2.1.B26 Saccharolobus solfataricus P2 P22498
-
-
3.2.1.B28 Pyrococcus furiosus Q51723
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.B26
-
 Saccharolobus solfataricus
3.2.1.B28
-
 Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.B26 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Saccharolobus solfataricus 4-nitrophenol + D-galactose
-
 ?
3.2.1.B26 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Saccharolobus solfataricus P2 4-nitrophenol + D-galactose
-
 ?
3.2.1.B26 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Saccharolobus solfataricus 4-nitrophenol + D-glucopyranose
-
 ?
3.2.1.B26 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Saccharolobus solfataricus P2 4-nitrophenol + D-glucopyranose
-
 ?
3.2.1.B26 cellobiose + H2O
-
 Saccharolobus solfataricus D-glucose + D-glucose
-
 ?
3.2.1.B26 cellobiose + H2O
-
 Saccharolobus solfataricus P2 D-glucose + D-glucose
-
 ?
3.2.1.B26 lactose + H2O
-
 Saccharolobus solfataricus D-glucose + D-galactose
-
 ?
3.2.1.B26 lactose + H2O
-
 Saccharolobus solfataricus P2 D-glucose + D-galactose
-
 ?
3.2.1.B28 4-nitrophenyl beta-D-galactopyranoside + H2O CelB is a little more active on the galactosides lactose and 4-nitrophenyl beta-D-galactopyranoside than on 4-nitrophenyl beta-D-glucopyranoside, whereas the efficiency of cellobiose hydrolysis is relatively low Pyrococcus furiosus 4-nitrophenol + D-galactose
-
 ?
3.2.1.B28 4-nitrophenyl beta-D-glucopyranoside + H2O CelB is a little more active on the galactosides lactose and 4-nitrophenyl beta-D-galactopyranoside than on 4-nitrophenyl beta-D-glucopyranoside, whereas the efficiency of cellobiose hydrolysis is relatively low Pyrococcus furiosus 4-nitrophenol + D-glucopyranose
-
 ?
3.2.1.B28 cellobiose + H2O CelB is a little more active on the galactosides lactose and 4-nitrophenyl beta-D-galactopyranoside than on 4-nitrophenyl beta-D-glucopyranoside, whereas the efficiency of cellobiose hydrolysis is relatively low Pyrococcus furiosus D-glucose + D-glucose
-
 ?
3.2.1.B28 lactose + H2O CelB is a little more active on the galactosides lactose and 4-nitrophenyl beta-D-galactopyranoside than on 4-nitrophenyl beta-D-glucopyranoside, whereas the efficiency of cellobiose hydrolysis is relatively low Pyrococcus furiosus D-glucose + D-galactose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.B26 LACS
-
 Saccharolobus solfataricus
3.2.1.B26 SSO3019 locus name Saccharolobus solfataricus
3.2.1.B28 CelB
-
 Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.B26 70
-
 assay at Saccharolobus solfataricus
3.2.1.B26 95
-
-
 Saccharolobus solfataricus
3.2.1.B28 70
-
 assay at Pyrococcus furiosus
3.2.1.B28 104
-
-
 Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.B26 70
-
 1.5 h, no residual activity Saccharolobus solfataricus
3.2.1.B26 92
-
 half-life: less than 3 min Saccharolobus solfataricus
3.2.1.B28 92
-
 half-life: above 100 min Pyrococcus furiosus
3.2.1.B28 104
-
 Tm-value Pyrococcus furiosus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.B26 23.2
-
 cellobiose pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 45
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 121
-
 lactose pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 165
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B28 37.6
-
 cellobiose pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 206
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 286
-
 lactose pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 298
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.0, 70°C Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.B26 5
-
 assay at Saccharolobus solfataricus
3.2.1.B28 5
-
 assay at Pyrococcus furiosus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.B26 0.0000084
-
 (5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol pH 5.0, 70°C, substrate: 4-nitrophenyl beta-D-galactopyranoside Saccharolobus solfataricus
3.2.1.B26 46
-
 D-glucose pH 5.0, 70°C, substrate: 4-nitrophenyl beta-D-galactopyranoside Saccharolobus solfataricus
3.2.1.B28 0.0000093
-
 (5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol pH 5.0, 70°C, substrate: 4-nitrophenyl beta-D-galactopyranoside Pyrococcus furiosus
3.2.1.B28 43
-
 D-glucose pH 5.0, 70°C, substrate: 4-nitrophenyl beta-D-galactopyranoside Pyrococcus furiosus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.B26 1.72
-
 cellobiose pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 3.19
-
 lactose pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 105
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B26 375
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.0, 70°C Saccharolobus solfataricus
3.2.1.B28 4.78
-
 lactose pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 11.8
-
 cellobiose pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 115
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.0, 70°C Pyrococcus furiosus
3.2.1.B28 258
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.0, 70°C Pyrococcus furiosus